Polypeptide having inhibitory effect on novel coronavirus HCoV-EMC/2012 infection and application thereof

A hcov-emc2012, coronavirus technology, applied in the field of recombinant cells, can solve the problem of no effective specific drugs for the new coronavirus HCoV-EMC, and achieve the effect of filling the international gap

Active Publication Date: 2014-10-01
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At the same time, there is no effective specific drug for the new coronavirus HCoV-EMC 2012...

Method used

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  • Polypeptide having inhibitory effect on novel coronavirus HCoV-EMC/2012 infection and application thereof
  • Polypeptide having inhibitory effect on novel coronavirus HCoV-EMC/2012 infection and application thereof
  • Polypeptide having inhibitory effect on novel coronavirus HCoV-EMC/2012 infection and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 The polypeptide enters the inhibition experiment on the new coronavirus HcoV-EMC 2012

[0039] The peptides HR1-1, HR1-2, HR1-3, HR1-4, HR1-5, HR2-1, HR2-2 and HR2-3 ( figure 1 Shown) Perform 4-fold gradient dilution; add 100 μl of each sample to a 96-well cell culture plate pre-plated with 50,000 African green monkey kidney cells (Vero cells) per well, incubate at 37°C for 2 hours, then remove the supernatant. Wash the cells twice with PBS, add 100 μl of new coronavirus HCoV-EMC 2012 (10TCID50), incubate at 37°C for 1 hour; then wash the cells twice with PBS. Then add 100 μl sample / well to the 96-well plate for further interaction with the cells. The cells were then cultured at 37°C for 3 days. Record the cytopathic effect (CPE), then add 10 μl MTT to each well. Incubate at 37°C for 4 hours; add 100 μl of 0.01M HCl containing 1% SDS to each well, incubate overnight at 37°C and measure the absorbance at a wavelength of 570nm.

[0040] Anti-virus results such as...

Embodiment 2

[0045] Example 2 Interaction between HR1-5 and HR2-2

[0046] This experiment refers to the document "Interaction between heptad repeat 1 and 2 regions in spike protein of SARS-associated coronavirus: implications for virus fusogenic mechanism and identification of fusion inhibitors." (Liu,SW et al. Lancet 363, 938-947, 2004) operating.

[0047]

[0048] 1. Non-denaturing polyacrylamide gel electrophoresis (N-PAGE). Mix HR1 series peptides with HR2 series peptides (final concentration 30μM each), and incubate at 37°C for 30 minutes. Take 5× high pH loading buffer and mix it with 4 times the volume of peptide mixture. Then spot the sample (25 μl per well) in 18% non-denaturing gel (Beijing Tianenze), and conduct electrophoresis at 125V constant voltage for 2 hours at room temperature. Coomassie brilliant blue stain. The results showed that after HR1-5 was mixed with HR2 series peptides, a new peptide was produced ( Figure 4 A). This band is the EMC virus fusion active six-hel...

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PUM

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Abstract

The invention belongs to the field of biological medicine and provides a polypeptide. The polypeptide contains a sequence as shown in SEQ ID No. 9 and has an inhibitory effect on novel coronavirus HCoV-EMC/2012 infection. The invention also provides to a nucleic acid molecule coding the polypeptide, related recombinant protein, a nucleic acid molecule, an expression vector and a recombinant cell. A medicine can be prepared from the polypeptide provided by the invention and frequently medically acceptable carrier, excipient or the like for prevention and inhibition of novel coronavirus HCoV-EMC/2012 infection; and polypeptide can also be combined with other medicines. The invention provides candidate polypeptide medicines for prevention or treatment of novel coronavirus HCoV-EMC/2012 infection.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to a group of polypeptides capable of inhibiting the infection of novel coronavirus HCoV-EMC 2012, as well as nucleic acid molecules encoding the polypeptides, related recombinant proteins, nucleic acid molecules, expression vectors and recombinant cells. Background technique [0002] In 2012, a new type of coronavirus HCoV-EMC 2012 originating from the Middle East has caused many deaths, and the mortality rate was as high as 57%, and the rest of the patients also suffered from severe diseases. Current studies have found that the receptor used by the new coronavirus HCoV-EMC 2012 is different from that used by the SARS coronavirus. Its receptor is dipeptidyl peptidase 4 (DPP4), a widely distributed human cell receptor, thus The virus has a wide range of infectivity to human cells. In addition, the new coronavirus HCoV-EMC 2012 can infect a variety of animals, including bats and pigs, makin...

Claims

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Application Information

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IPC IPC(8): C07K14/165C12N15/50C12N15/63A61K39/215A61K48/00A61P31/14C12N5/071
CPCA61K38/00A61K48/00C12N15/63C07K14/165A61K39/215C07K14/005C12N2770/20022C12N2770/20033A61P31/14
Inventor 姜世勃陆路刘奇
Owner FUDAN UNIV
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